YM58483 是一种特异性有效的 CRAC 通道和随后的 Ca2+ 信号抑制剂,可阻断钙池操纵的阳离子内流 (SOCE)。
产品描述
YM-58483 is a specific and effective inhibitor of CRAC channels and subsequent Ca2+ signals.
体外活性
在多种过敏性哮喘模型,如气道高反应、早期和晚期支气管狭窄、抗原诱导的气道嗜酸性粒细胞中,YM-58483均有抑制效果,在大鼠和豚鼠器官中还出现白三烯、IL-4水平的降低.在GVHD小鼠中,YM-58483还可抑制抗宿主CTL反应、供体T细胞扩张及IFN-γ的产生.YM-58483(30 mg/kg,p.o.)对小鼠的一般活动没有明显影响[1].
体内活性
作为选择性SOCE阻滞剂,YM-58483在Jurkat T细胞中抑制anti-CD3抗体所诱导的持续性钙离子流入。它抑制CRAC、TRPC3和TRPC5通道、促进TRPM4通道,抑制细胞因子的产生(IL-2, IL-4, IL-5, IFN-γ等)及T细胞的增殖。通过抑制NF-AT的激活,YM-58483可抑制MLR相关的脾细胞增殖[1]。YM-58483对IL-2的产生和NF-AT驱动的启动子活性也有明显的抑制作用,但不影响Jurkat细胞中AP-1驱动的启动子活性[2]。
激酶实验
HCT-116 cells are washed with PBS and then homogenized with a 27-gauge syringe in binding buffer (10 mm Tris-HCl (pH 7.4), 50 mm KCl, 5 mm MgCl2, 1 mm EDTA, and 0.1 mm Na3VO4). The cell lysate is centrifuged at 13,000 rpm for 30 min at 4°C, and the supernatant is collected. The HCT-116 cell lysate supernatant is precleared by incubating with Dynabeads M-280 streptavidin for 30 min at 4°C and captured by magnet separation. The cleared supernatants are incubated with biotinyl-KRIBB11 compound. After overnight incubation at 4°C, proteins associated with the biotinyl-KRIBB11 compound are precipitated with Dynabeads M-280 streptavidin. Precipitated samples are separated by a magnet. Samples are washed with 1 mL of ishing buffer containing 50 mm HEPES (pH 7.5), 50 mm NaCl, 1 mm EDTA, 1 mm EGTA, 0.1% Tween 20, 10% (v/v) glycerol, 1 mm NaF, 0.1 mm Na3VO4, and protease inhibitor mixture tablets (1 tablet/10 mL). Samples are boiled in SDS-PAGE sample buffer, separated by 10% polyacrylamide gel, and immunoblotted with antibodies against HSF1, HSF2, HSP90, or CDK9.
细胞实验
Jurkat cells (1×107 cells/ml) were tested with varying concentration of compounds for 30 min at 37°C. The cells were stimulated with 1 μM ionomycin for 30 min at 37°C. After stimulation, the cells were centrifuged at 200×g for 2 min, and were solubilized in 100 μl of Triton X-100 lysis buffer. The cell lysate was centrifuged at 15,000×g for 20 min; the clarified lysate was subjected to SDS-PAGE; and NF-ATc2 was detected by Western blotting with anti-NF-ATc2 mAb. (Only for Reference)
Cas No.
223499-30-7
分子式
C15H9F6N5OS
分子量
421.32
别名
BTP2;YM 58483
储存和溶解度
Ethanol:78 mg/mL (185.1 mM)
H2O:<1 mgml
DMSO:78 mg/mL (185.1 mM)
Powder: -20°C for 3 years
In solvent: -80°C for 2 years